La técnica quirúrgica fue adquirido originalmente en el laboratorio del doctor William D. Hill, en el Colegio Médico de Georgia. Los autores también desean agradecer al Dr. David A. Rempe y Prifti Landa para el uso de la cámara de disección. Esta investigación fue financiada por el NIH NS041744, NS051279, NS064700 F31 y 30815697D AHA.

Este protocolo fue aprobado por la Universidad de Rochester comité dedicado a la ética del uso de animales en la investigación (UCAR). Técnicas de asepsia deben ser seguidas durante el protocolo. El uso de guantes estériles y mascarilla es necesario. Todos los equipos, materiales, productos químicos y herramientas que se utilizan en el protocolo se describen en la Tabla 1. 1. La preparación pre-quirúrgica <ol><li> Se inyecta por vía subcutánea ratones con buprenorfina (0.05mg/kg) 2 h antes de la cirugía, inmediatamente después de la cirugía y luego cada 3-5 h durante las primeras 24 horas post-operatorio. </li><li> Esterilizar todos los instrumentos quirúrgicos, gasas, algodón y aplicadores de punta por autoclave. Mantenga las herramientas quirúrgicas en el 70% de etanol durante la cirugía y el aire seco en una gasa estéril justo antes de su uso. Rocíe el área de trabajo con el 70% de etanol. </li><li> Anestesiar el ratón con un 3% isofluorano-20% mezcla de gas de oxígeno mediante el uso de un vaporizador anestésico. Ajustar la cantidad de oxígeno con el medidor de flujo. Evaluar el nivel de anestesia por los pies o los pellizcos de cola (que debe ser que no responde). Mantener el nivel de la anestesia con un 2% (v / v) isofluorano. </li><li> Aplicar lágrimas artificiales para los ojos del ratón y tenga cuidado para evitar daños en el ojo durante la intervención quirúrgica. Coloque el ratón sobre su lado derecho en posición lateral. </li><li> Afeitar el área en el lado izquierdo entre el ojo izquierdo y la base de la oreja izquierda con los Clippers de los animales. Limpie el área por la alternancia entre una solución de Betadine y el 70% de etanol con aplicadores con punta de algodón y frotar suavemente la zona. Repita según sea necesario. </li><li> Coloque el ratón sobre una almohadilla eléctrica conectada a una sonda rectal para mantener la temperatura corporal a 37 º C. Inserte la sonda rectal mediante el uso de aceite mineral. </li><li> Coloque el ratón sobre su lado derecho bajo el microscopio y seguro con cinta adhesiva. Cortar una ventana en una gasa y cubrir el área quirúrgica. </li></ol> 2. Procedimiento quirúrgico y la ligadura de MCA <ol><li> Haga una incisión vertical entre el ojo izquierdo y la base de la oreja izquierda con finas tijeras rectas. Use pinzas hemostáticas curvas para mantener el área de cirugía abierta. </li><li> Se practica una incisión horizontal en el músculo temporal mediante el uso de tijeras de primavera y un poco separado del músculo temporal del cráneo tirando lentamente con unas pinzas. </li><li> Hacer una craneotomía subtemporal pequeña en la unión del arco cigomático y hueso escamoso mediante el uso de una aguja 18G, mientras que la celebración de la mandíbula con pinzas curvas. </li><li> Exponer la MCA mediante la eliminación de los pequeños trozos de cráneo con gubia ósea. El arco cigomático y contenido de la órbita no debe ser dañada durante el proceso. Si el secado excesivo de los tejidos se produce durante este proceso, se aplican PBS estéril con aplicadores con punta de algodón. </li><li> Ligar la porción distal de la MCA mediante el uso de un cauterizador de pequeños vasos. </li><li> Lugar del músculo temporal en su posición original y cerrar el sitio de la incisión quirúrgica con suturas de nylon 5-0. </li><li> Deje de anestesia y retire la sonda rectal. Inyectar el ratón con una 2 ª dosis de buprenorfina (0.05mg/kg) por vía subcutánea. Devolver el ratón de la jaula que se mantuvo a 37 ° C con un panel de calefacción. </li><li> Vigilar de cerca el ratón durante las próximas 24 horas para cualquier molestia (disminución del apetito / consumo de agua, postura encorvada, aumento de la respiración, pilo-erigido pelo). Inyectar el ratón con la buprenorfina por vía subcutánea cada 3-5 h después de la cirugía hasta 24 horas. Proporcionar el ratón con gel de recuperación durante este período. </li></ol> 3. TTC tinción y determinación del volumen sistólico <ol><li> Veinticuatro horas después de la cirugía profundamente anestesiar el ratón, el ratón transcardially perfundir con un 4% IVA incluido (w / v) en tampón fosfato salino (PBS) durante 15 minutos y luego con una solución de paraformaldehído al 4% durante 10 minutos usando una mini-bomba peristáltica a un flujo medio. Quitar el cerebro y colocarlo en la solución de paraformaldehído al 4% durante la noche. </li><li> La posición del cerebro en el bloque de corte. Cortar las rebanadas de cerebro en 1 mm de espesor mediante el uso de hojas de afeitar. </li><li> Colocar las rebanadas de lado de una regla escala milimétrica. Fotografía de los cortes mediante el uso de una cámara digital conectada al microscopio de disección. </li><li> Calcular el área de derrame cerebral al restar el área no infartada del sitio ipsilateral de la superficie total del sitio contralateral usando la imagen J Software (http://rsbweb.nih.gov/ij/). Calcular el volumen de movimiento de la estiba del área de la apoplejía de los 22 cortes. </li></ol> 4. Instrucciones para utilizar el software Image J <ol><li> Abra el archivo de imagen para ser analizadas en el software Image J haciendo clic en el menú de archivo. </li><li> Haga clic en la pestaña &quot;línea recta&quot; en el programa. Trace una línea recta entre los dos márgenes de la regla. Haga clic en &quot;analizar&quot; y seleccione &quot;escala de conjunto&quot;. Establecer la distancia conocida como 1, unidad de longitud en mm. </li><li> Haga clic en la pestaña &quot;círculo a mano alzada. Dibuje un círculo para delinear el hemisferio contralateral. Haga clic en &quot;analizar&quot; y seleccione &quot;medida&quot;. La ventana de cálculo se abrirá mostrando el área del círculo dibujado. </li><li> Dibuja otro círculo alrededor del hemisferio ipsilateral exclusión de accidente cerebrovascular (blanco) la zona. Medir el área, como se indica en el paso 4.3. El nuevo valor se añade a la ventana de cálculo. La diferencia entre la primera y la segunda representa los valores de la zona del infarto que se indica como A en la fórmula a continuación. </li><li> Calcular el área del accidente cerebrovascular en cada rebanada repitiendo los pasos 4.2-4.4. Tome la suma del área de movimiento calculado en cada sector (ΣA n). Esto representa el volumen de eyección conocer el hecho de que el grosor de cada rebanada es 1 mm. ΣA n x 1 mm (grosor de cada rebanada) = volumen Carrera </li></ol> 5. Los resultados representativos: Los infartos obtenidos por la ligadura permanente de la MCA en el ratón son en su mayoría cortical. Sin embargo, es posible obtener lesiones subcorticales si el MCA se liga proximal a la rama lenticuloestriadas. Los volúmenes de derrame cerebral después de la ligadura de MCA puede variar de 10 mm a 35 mm ³ ³ 19, 23. El volumen sistólico calculado con la tinción de TTC en Moyanova et al. 19 se encuentran entre ³ 10 mm a 22 mm ³, mientras que el volumen sistólico determina a partir de imágenes de resonancia magnética en Filiano et al 23 se encuentran entre ³ 20 mm a 35 mm ³. La posible razón de estas diferencias tal vez la ubicación exacta de la ligadura y los diferentes métodos utilizados para medir el volumen sistólico. En la Figura 1, un manchado TTC cerebral 24 horas después de la ligadura permanente MCA en una de tipo salvaje C57/BL6 del ratón se muestra. El sitio de accidente cerebrovascular es de apariencia blanca (Fig. 1A, B). Figura 1 A y B ilustran el sitio del infarto desde diferentes ángulos. La figura 2 muestra rodajas de 1 mm de espesor de la TTC manchada cerebro tiempos de adelante hacia atrás (Fig. 2A-G). El volumen del infarto se calculó 24 horas después de la cirugía. El volumen del infarto es ³ ~ 23mm. <img src="/files/ftp_upload/3039/3039fig1.jpg" alt="Figura 1"/> Figura 1. Representación de sitio accidente cerebrovascular. AB, TTC manchadas imágenes a todo el cerebro, 24 horas después de la ligadura de MCA. Área blanca representa el infarto. <img src="/files/ftp_upload/3039/3039fig2.jpg" alt="Figura 2"/> Figura 2. Los resultados representativos. (A, G&gt;), IVA incluido manchado secciones de cerebro de 1 mm, 24 horas después de MCAL. Los sectores se alinean de adelante hacia atrás. Área blanca representa el infarto.

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El método permanente MCA ligadura da volumen del infarto altamente reproducible y el aumento de post-operatorio las tasas de supervivencia en comparación con otros métodos disponibles. La facilidad y la corta duración (~ 30 min) del procedimiento que sea aún más práctico. El método es ampliamente utilizado tanto en ratones y ratas. Esta técnica requiere una cirugía invasiva bajo un microscopio estereoscópico. Por lo tanto, la experiencia en la operación bajo el microscopio y el p...

<table border="1"><tbody><tr><th> NOMBRE </th><th> Empresa y el número CATÁLOGO </th></tr><tr><td> A. Soluciones y Productos Químicos </td><td></td></tr><tr><td> Solución de povidona yodada, el lavado quirúrgico </td><td> Aplicare, 82-209 </td></tr><tr><td> 70% de etanol </td><td> Koptec, V1001 </td></tr><tr><td> 2,3,5 cloruro de trifenil tetrazolio (TTC) </td><td> Sigma, T8877 </td></tr><tr><td> Paraformaldehído al 4% </td><td> Microscopía electrónica de Ciencias, 159SP </td></tr><tr><td> Tampón fosfato pH Saline,: 7.4 </td><td> Hecho y se esterilizan en el laboratorio. </td></tr><tr><td> Aceite mineral </td><td> PML microbiológicos, R6570 </td></tr><tr><td> Isofluorano </td><td> La Compañía Butler, 029405 </td></tr><tr><td> La buprenorfina (0.3mg/ml) </td><td> Hospira, NDC 0409-2012-32 </td></tr><tr><td> Las lágrimas artificiales </td><td> La Compañía Butler, 007312 </td></tr><tr><td> Recuperación de Gel </td><td> Claro H 2 O, 72-01-5022 </td></tr><tr><td> B. Materiales y equipo quirúrgico </td><td></td></tr><tr><td> C57/BL6 ratón, 10-12 semanas de edad </td><td> La colonia fue criado en el vivero universitario </td></tr><tr><td> 100% tanque de oxígeno </td><td> Airgas </td></tr><tr><td> Vaporizador anestésico y un medidor de flujo </td><td> Surgivet, Modelo 100 </td></tr><tr><td> Almohadilla eléctrica con sonda rectal </td><td> Herramientas de Bellas Ciencia, TR200 </td></tr><tr><td> Microscopio de disección </td><td> Zeiss, Stemi 2000 </td></tr><tr><td> Calefacción de </td><td> Petco Services Ltd. 0307-013 </td></tr><tr><td> Embarcación pequeña Set cauterizador </td><td> Herramientas de Bellas Ciencia, 18000-00 </td></tr><tr><td> Mini bomba peristáltica </td><td> Harvard Apparatus, GP II </td></tr><tr><td> 1 ml jeringa </td><td> BD, 329650 </td></tr><tr><td> 18G aguja </td><td> Becton Dickinson, 305196 </td></tr><tr><td> Quirúrgico 5,0 sutura de nylon </td><td> Ethilon, 698H </td></tr><tr><td> Bloque de seccionamiento </td><td> Kent Científico, 1mm </td></tr><tr><td> Hojas de un solo filo de la navaja </td><td> Microscopía electrónica de Ciencias, 71962 </td></tr><tr><td> Etiqueta de la cinta </td><td> Scienceware, Bel-Art Productos F13463-2075 </td></tr><tr><td> Esponjas de gasa </td><td> Kendall Curity 2252, Johnson y Johnson 6415 </td></tr><tr><td> Aplicadores con punta de algodón </td><td> Puritana 806-CMT </td></tr><tr><td> C. Cirugía Herramientas </td><td></td></tr><tr><td> Tijeras quirúrgicas primavera </td><td> Herramientas de Bellas Ciencia 15003-08 </td></tr><tr><td> Pinza en ángulo Microdissecting </td><td> Herramientas de Bellas Ciencia 5 / 45 </td></tr><tr><td> Pinzas curvas Microdissecting </td><td> Bellas Ciencia Herramientas 5 Ti </td></tr><tr><td> Bellas tijeras rectas </td><td> Herramientas de Bellas Ciencia 14568-12 </td></tr><tr><td> Gubia hueso curvo </td><td> Herramientas de Bellas Ciencia 16021-14 </td></tr><tr><td> Pinzas hemostáticas curvas Hartman </td><td> Herramientas de Bellas Ciencia 13003-10 </td></tr></tbody></table> Tabla 1. La lista de materiales, herramientas y productos químicos utilizados para el procedimiento de ligadura de MCA. A. Las soluciones y productos químicos, materiales quirúrgicos y B. Equipos, Herramientas C. Cirugía.

the application of permanent middle cerebral artery ligation in the mouse

Isquemia cerebral focal se encuentra entre el tipo más común de accidente cerebrovascular en los pacientes. Debido a la importancia clínica no ha habido un esfuerzo prolongado para desarrollar modelos animales para estudiar los acontecimientos que se desarrollan durante la lesión isquémica. Estas técnicas incluyen transitoria o permanente, los modelos de isquemia focal o global con muchos diferentes modelos animales, con los roedores más comunes son. El método permanente MCA ligadura que también se conoce como pMCAo en la literatura se utiliza ampliamente como un modelo de isquemia focal en los roedores 1-6. Este método fue descrito originalmente para las ratas por Tamura et al. en 1981 7. En este protocolo se utilizó una craneotomía para acceder a la MCA y de las regiones proximal se ocluye por electrocoagulación. Los infartos a regiones corticales y sobre todo del estriado a veces dependiendo de la localización de la oclusión. Esta técnica está bien establecido y utilizado en muchos laboratorios de 8.13. El uso precoz de esta técnica condujo a la definición y descripción del &quot;núcleo del infarto&quot; y &quot;penumbra&quot;, 14-16, y es a menudo utilizado para evaluar el potencial de compuestos neuroprotectores 10, 12, 13, 17. Aunque los estudios iniciales se realizaron en ratas, la ligadura permanente MCA ha sido utilizado con éxito en ratones con ligeras modificaciones, 18-20. Este modelo produce infartos reproducible y el aumento de las tasas de supervivencia después. Aproximadamente el 80% de los accidentes cerebrovasculares isquémicos en los humanos ocurren en el área de MCA 21 y por lo tanto este es un modelo muy importante para los estudios de tiempos. En la actualidad, hay una escasez de tratamientos eficaces para los pacientes con ictus, y por lo tanto hay una necesidad de un buen modelo para evaluar el potencial compuestos farmacológicos y evaluar los resultados fisiológicos. Este método también puede ser utilizado para el estudio de los mecanismos de respuesta a la hipoxia intracelular in vivo. A continuación, presentamos la cirugía de ligadura de MCA en un ratón C57/BL6. Se describe la preparación pre-quirúrgica, la cirugía de ligadura de MCA y 2,3,5 cloruro de trifenil tetrazolio (TTC) tinción para la cuantificación de los volúmenes de infarto.

Watch this Scientific Journal Video about The Application Of Permanent Middle Cerebral Artery Ligation in the Mouse at JoVE.com

The Application Of Permanent Middle Cerebral Artery Ligation in the Mouse

La arteria cerebral media (MCA) ligadura es una técnica para estudiar la isquemia cerebral focal en modelos animales. En este método, la arteria cerebral media se expone mediante craneotomía y se liga con cauterización. Este método da volumen del infarto altamente reproducible y el aumento de post-operatorio las tasas de supervivencia en comparación con otros métodos disponibles.

La aplicación de la Permanente Ligadura arteria cerebral media en el ratón

Focal cerebral ischemia is among the most common type of stroke seen in patients. Due to the clinical significance there has been a prolonged effort to ...

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Research

JoVE Journal

Medicine

19.5K vistas.  University of Rochester. La arteria cerebral media (MCA) ligadura es una técnica para estudiar la isquemia cerebral focal en modelos animales. En este método, la arteria cerebral media se expone mediante craneotomía y se liga con cauterización. Este método da volumen del infarto altamente reproducible y el aumento de post-operatorio las tasas de supervivencia en comparación con otros métodos disponibles.

Video: The Application Of Permanent Middle Cerebral Artery Ligation in the Mouse

Modeling Stroke in Mice - Middle Cerebral Artery Occlusion with the Filament Model

Stroke is among the most frequent causes of death and adult disability, especially in highly developed countries. However, treatment options to date are very limited. To meet the need for novel therapeutic approaches, experimental stroke research frequently employs rodent models of focal cerebral ischaemia. Most researchers use permanent or transient occlusion of the middle cerebral artery (MCA) in mice or rats.
Proximal occlusion of the middle cerebral artery (MCA) via the intraluminal suture technique (so called filament or suture model) is probably the most frequently used model in experimental stroke research. The intraluminal MCAO model offers the advantage of inducing reproducible transient or permanent ischaemia of the MCA territory in a relatively non-invasive manner. Intraluminal approaches interrupt the blood flow of the entire territory of this artery. Filament occlusion thus arrests flow proximal to the lenticulo-striate arteries, which supply the basal ganglia. Filament occlusion of the MCA results in reproducible lesions in the cortex and striatum and can be either permanent or transient. In contrast, models inducing distal (to the branching of the lenticulo-striate arteries) MCA occlusion typically spare the striatum and primarily involve the neocortex. In addition these models do require craniectomy. In the model demonstrated in this article, a silicon coated filament is introduced into the common carotid artery and advanced along the internal carotid artery into the Circle of Willis, where it blocks the origin of the middle cerebral artery. In patients, occlusions of the middle cerebral artery are among the most common causes of ischaemic stroke. Since varying ischemic intervals can be chosen freely in this model depending on the time point of reperfusion, ischaemic lesions with varying degrees of severity can be produced. Reperfusion by removal of the occluding filament at least partially models the restoration of blood flow after spontaneous or therapeutic (tPA) lysis of a thromboembolic clot in humans.
In this video we will present the basic technique as well as the major pitfalls and confounders which may limit the predictive value of this model.

Filamentous occlusion of the Middle cerebral artery is a common model for studying ischemic stroke in mice.

Modelado de accidente cerebrovascular en ratones - Oclusión arteria cerebral media con el modelo de filamento

Stroke is among the most frequent causes of death and adult disability, especially in highly developed countries. However, treatment options to date are ...

Investigación

Medicina

Mouse Model of Middle Cerebral Artery Occlusion

Stroke is the most common fatal neurological disease in the United States 1. The majority of strokes (88%) result from blockage of blood vessels in the brain (ischemic stroke) 2. Since most ischemic strokes (~80%) occur in the territory of middle cerebral artery (MCA) 3, many animal stroke models that have been developed have focused on this artery. The intraluminal monofilament model of middle cerebral artery occlusion (MCAO) involves the insertion of a surgical filament into the external carotid artery and threading it forward into the internal carotid artery (ICA) until the tip occludes the origin of the MCA, resulting in a cessation of blood flow and subsequent brain infarction in the MCA territory 4. The technique can be used to model permanent or transient occlusion 5. If the suture is removed after a certain interval (30 min, 1 h, or 2 h), reperfusion is achieved (transient MCAO); if the filament is left in place (24 h) the procedure is suitable as a model of permanent MCAO. This technique does not require craniectomy, a neurosurgical procedure to remove a portion of skull, which may affect intracranial pressure and temperature 6. It has become the most frequently used method to mimic permanent and transient focal cerebral ischemia in rats and mice 7,8. To evaluate the extent of cerebral infarction, we stain brain slices with 2,3,5-triphenyltetrazolium chloride (TTC) to identify ischemic brain tissue 9. In this video, we demonstrate the MCAO method and the determination of infarct size by TTC staining.

We demonstrate in the video a method for producing a middle cerebral artery occlusion in adult mice using an intraluminal monofilament. We also show how to evaluate the extent of cerebral infarction by 2,3,5-triphenyltetrazolium chloride (TTC) staining.

Modelo de ratón de la oclusión arteria cerebral media

Stroke is the most common fatal neurological disease in the United States 1. The majority of strokes (88%) result from blockage of blood vessels in the ...

Utilizing a Cranial Window to Visualize the Middle Cerebral Artery During Endothelin-1 Induced Middle Cerebral Artery Occlusion

Creation of a cranial window is a method that allows direct visualization of structures on the cortical surface of the brain1-3. This technique can be performed in many locations overlying the rat cerebrum, but is most easily carried out by creating a craniectomy over the readily accessible frontal or parietal bones. Most frequently, we have used this technique in combination with the endothelin-1 middle cerebral artery occlusion model of ischemic stroke to quantify the changes in middle cerebral artery vessel diameter that occur with injection of endothelin-1 into the brain parenchyma adjacent to the proximal MCA4, 5. In order to visualize the proximal portion of the MCA during endothelin -1 induced MCAO, we use a technique to create a cranial window through the temporal bone on the lateral aspect of the rat skull (Figure 1). Cerebral arteries can be visualized either with the dura intact or with the dura incised and retracted. Most commonly, we leave the dura intact during visualization since endothelin-1 induced MCAO involves delivery of the vasoconstricting peptide into the brain parenchyma. This bypasses the need to incise the dura directly over the visualized vessels for drug delivery. This protocol will describe how to create a cranial window to visualize cerebral arteries in a step-wise fashion, as well as how to avoid many of the potential pitfalls pertaining to this method.

This article describes a method for visualizing rat cerebral arteries through a cranial window using temporal craniectomy in order to view proximal portions of the middle cerebral artery (Figure 1). This versatile method can be combined with various techniques of drug delivery to measure cerebral artery reactivity in vivo.

Utilizando una ventana craneal para visualizar la arteria cerebral media Durante la endotelina-1 inducida por oclusión de la arteria cerebral media

Creation of a cranial window is a method that allows direct visualization of structures on the cortical surface of the brain1-3. This technique can be ...

Optimized System for Cerebral Perfusion Monitoring in the Rat Stroke Model of Intraluminal Middle Cerebral Artery Occlusion

The translational potential of pre-clinical stroke research depends on the accuracy of experimental modeling. Cerebral perfusion monitoring in animal models of acute ischemic stroke allows to confirm successful arterial occlusion and exclude subarachnoid hemorrhage. Cerebral perfusion monitoring can also be used to study intracranial collateral circulation, which is emerging as a powerful determinant of stroke outcome and a possible therapeutic target. Despite a recognized role of Laser Doppler perfusion monitoring as part of the current guidelines for experimental cerebral ischemia, a number of technical difficulties exist that limit its widespread use. One of the major issues is obtaining a secure and prolonged attachment of a deep-penetration Laser Doppler probe to the animal skull. In this video, we show our optimized system for cerebral perfusion monitoring during transient middle cerebral artery occlusion by intraluminal filament in the rat. We developed in-house a simple method to obtain a custom made holder for twin-fibre (deep-penetration) Laser Doppler probes, which allow multi-site monitoring if needed. A continuous and prolonged monitoring of cerebral perfusion could easily be obtained over the intact skull.

Cerebral perfusion monitoring has been demonstrated to improve accuracy in ischemic stroke models. Technical difficulties often limit the use of this essential tool for cerebrovascular research. In this video, an optimized system is shown to obtain a single or multi-site hemodynamic monitoring during intraluminal middle cerebral artery occlusion in rats.

Sistema optimizado para la monitorización de la perfusión cerebral en el modelo de rata de Carrera intraluminal oclusión arteria cerebral media

The translational potential of pre-clinical stroke research depends on the accuracy of experimental modeling. Cerebral perfusion monitoring in animal ...

Permanent Cerebral Vessel Occlusion via Double Ligature and Transection

Stroke is a leading cause of death, disability, and socioeconomic loss worldwide. The majority of all strokes result from an interruption in blood flow (ischemia) 1. Middle cerebral artery (MCA) delivers a great majority of blood to the lateral surface of the cortex 2, is the most common site of human stroke 3, and ischemia within its territory can result in extensive dysfunction or death 1,4,5. Survivors of ischemic stroke often suffer loss or disruption of motor capabilities, sensory deficits, and infarct. In an effort to capture these key characteristics of stroke, and thereby develop effective treatment, a great deal of emphasis is placed upon animal models of ischemia in MCA.
Here we present a method of permanently occluding a cortical surface blood vessel. We will present this method using an example of a relevant vessel occlusion that models the most common type, location, and outcome of human stroke, permanent middle cerebral artery occlusion (pMCAO). In this model, we surgically expose MCA in the adult rat and subsequently occlude via double ligature and transection of the vessel. This pMCAO blocks the proximal cortical branch of MCA, causing ischemia in all of MCA cortical territory, a large portion of the cortex. This method of occlusion can also be used to occlude more distal portions of cortical vessels in order to achieve more focal ischemia targeting a smaller region of cortex. The primary disadvantages of pMCAO are that the surgical procedure is somewhat invasive as a small craniotomy is required to access MCA, though this results in minimal tissue damage. The primary advantages of this model, however, are: the site of occlusion is well defined, the degree of blood flow reduction is consistent, functional and neurological impairment occurs rapidly, infarct size is consistent, and the high rate of survival allows for long-term chronic assessment.

Permanent Cerebral Vessel Occlusion via Double Ligature and Transection

We describe a highly reproducible method for the permanent occlusion of a rodent major cerebral blood vessel. This technique can be accomplished with very little peripheral damage, minimal blood loss, a high rate of long-term survival, and consistent infarct volume commensurate with the human clinical population.

Permanente oclusión vascular cerebral<em&gt; Vía</em&gt; Ligadura doble y transección

Stroke is a leading cause of death, disability, and socioeconomic loss worldwide. The majority of all strokes result from an interruption in blood flow ...

Modeling Stroke in Mice: Permanent Coagulation of the Distal Middle Cerebral Artery

Stroke is the third most common cause of death and a main cause of acquired adult disability in developed countries. Only very limited therapeutical options are available for a small proportion of stroke patients in the acute phase. Current research is intensively searching for novel therapeutic strategies and is increasingly focusing on the sub-acute and chronic phase after stroke because more patients might be eligible for therapeutic interventions in a prolonged time window. These delayed mechanisms include important pathophysiological pathways such as post-stroke inflammation, angiogenesis, neuronal plasticity and regeneration. In order to analyze these mechanisms and to subsequently evaluate novel drug targets, experimental stroke models with clinical relevance, low mortality and high reproducibility are sought after. Moreover, mice are the smallest mammals in which a focal stroke lesion can be induced and for which a broad spectrum of transgenic models are available. Therefore, we describe here the mouse model of transcranial, permanent coagulation of the middle cerebral artery via electrocoagulation distal of the lenticulostriatal arteries, the so-called &#8220;coagulation model&#8221;. The resulting infarct in this model is located mainly in the cortex; the relative infarct volume in relation to brain size corresponds to the majority of human strokes. Moreover, the model fulfills the above-mentioned criteria of reproducibility and low mortality. In this video we demonstrate the surgical methods of stroke induction in the &#8220;coagulation model&#8221; and report histological and functional analysis tools.

Various murine models of middle cerebral artery occlusion (MCAO) are widely used in experimental brain research. Here, we demonstrate the model of transcranial permanent distal MCAO which produces consistent cortical infarction of a size corresponding to damage imposed by the majority of human ischemic strokes.

Carrera Modelado en ratones: Coagulación permanente de la arteria cerebral media Distal

Stroke is the third most common cause of death and a main cause of acquired adult disability in developed countries. Only very limited therapeutical ...

Permanent Ligation of the Left Anterior Descending Coronary Artery in Mice: A Model of Post-myocardial Infarction Remodelling and Heart Failure

Heart failure is a syndrome in which the heart fails to pump blood at a rate commensurate with cellular oxygen requirements at rest or during stress. It is characterized by fluid retention, shortness of breath, and fatigue, in particular on exertion. Heart failure is a growing public health problem, the leading cause of hospitalization, and a major cause of mortality. Ischemic heart disease is the main cause of heart failure.
Ventricular remodelling refers to changes in structure, size, and shape of the left ventricle. This architectural remodelling of the left ventricle is induced by injury (e.g., myocardial infarction), by pressure overload (e.g., systemic arterial hypertension or aortic stenosis), or by volume overload. Since ventricular remodelling affects wall stress, it has a profound impact on cardiac function and on the development of heart failure. A model of permanent ligation of the left anterior descending coronary artery in mice is used to investigate ventricular remodelling and cardiac function post-myocardial infarction. This model is fundamentally different in terms of objectives and pathophysiological relevance compared to the model of transient ligation of the left anterior descending coronary artery. In this latter model of ischemia/reperfusion injury, the initial extent of the infarct may be modulated by factors that affect myocardial salvage following reperfusion. In contrast, the infarct area at 24 hr after permanent ligation of the left anterior descending coronary artery is fixed. Cardiac function in this model will be affected by 1) the process of infarct expansion, infarct healing, and scar formation; and 2) the concomitant development of left ventricular dilatation, cardiac hypertrophy, and ventricular remodelling.
Besides the model of permanent ligation of the left anterior descending coronary artery, the technique of invasive hemodynamic measurements in mice is presented in detail.

Heart failure is the leading cause of hospitalization and a major cause of mortality. A model of permanent ligation of the left anterior descending coronary artery in mice is applied to investigate ventricular remodelling and cardiac dysfunction post-myocardial infarction. The technique of invasive hemodynamic measurements in mice is presented.

La ligadura permanente de la arteria coronaria descendente anterior en ratones: un modelo de remodelación post infarto de miocardio e insuficiencia cardiaca

Heart failure is a syndrome in which the heart fails to pump blood at a rate commensurate with cellular oxygen requirements at rest or during stress. It ...

Performing Permanent Distal Middle Cerebral with Common Carotid Artery Occlusion in Aged Rats to Study Cortical Ischemia with Sustained Disability

Stroke typically occurs in elderly people with a range of comorbidities including carotid (or other arterial) atherosclerosis, high blood pressure, obesity and diabetes. Accordingly, when evaluating therapies for stroke in animals, it is important to select a model with excellent face validity. Ischemic stroke accounts for 80% of all strokes, and the majority of these occur in the territory of the middle cerebral artery (MCA), often inducing infarcts that affect the sensorimotor cortex, causing persistent plegia or paresis on the contralateral side of the body. We demonstrate in this video a method for producing ischemic stroke in elderly rats, which causes sustained sensorimotor disability and substantial cortical infarcts. Specifically, we induce permanent distal middle cerebral artery occlusion (MCAO) in elderly female rats by using diathermy forceps to occlude a short segment of this artery. The carotid artery on the ipsilateral side to the lesion was then permanently occluded and the contralateral carotid artery was transiently occluded for 60 min. We measure the infarct size using structural T2-weighted magnetic resonance imaging (MRI) at 24 hr and 8 weeks after stroke. In this study, the mean infarct volume was 4.5% &#177; 2.0% (standard deviation) of the ipsilateral hemisphere at 24 hr (corrected for brain swelling using Gerriet&#8217;s equation, n = 5). This model is feasible and clinically relevant as it permits the induction of sustained sensorimotor deficits, which is important for the elucidation of pathophysiological mechanisms and novel treatments.

Here we present a protocol to produce permanent distal middle cerebral artery occlusion in elderly female rats with simultaneous occlusion of the carotid arteries to generate large cortical infarcts and sustained deficits. We show confirmation of the lesion size using structural MRI at 24 hr and 8 weeks after stroke.

Realización Permanente distal cerebral media carótida común con oclusión de la arteria en ratas de edad avanzada para el Estudio de la isquemia cortical con sostenida de la discapacidad

Stroke typically occurs in elderly people with a range of comorbidities including carotid (or other arterial) atherosclerosis, high blood pressure, ...

Transient Middle Cerebral Artery Occlusion Model of Neonatal Stroke in P10 Rats

A number of animal models have been used to study hypoxic-ischemic injury, traumatic injury, global hypoxia, or permanent ischemia in both the immature and mature brain. Stroke occurs commonly in the perinatal period in humans, and transient ischemia-reperfusion is the most common form of stroke in neonates. The reperfusion phase is a critical component of injury progression, which occurs over a period of days to weeks, and of the endogenous response to injury. This postnatal day 10 (p10) rat model of transient middle cerebral artery occlusion (tMCAO) creates a unilateral, non-hemorrhagic focal ischemia-reperfusion injury that can be utilized to study the mechanisms of focal injury and repair in the full-term-equivalent brain. The injury pattern that is produced by tMCAO is consistent and highly reproducible and can be confirmed with MRI or histological analyses. The severity of injury can be manipulated through changes in occlusion time and other methods that will be discussed.

Neonatal stroke is a significant cause of early brain injury requiring a translational model with consistent focal injury patterns and high reproducibility in order to enable study. This study describes the detailed surgical procedure for creating a non-hemorrhagic, unilateral focal ischemia-reperfusion injury in full-term-equivalent rodents.

Transitoria de la arteria cerebral media oclusión Modelo de accidente cerebrovascular neonatal en ratas P10

A number of animal models have been used to study hypoxic-ischemic injury, traumatic injury, global hypoxia, or permanent ischemia in both the immature ...

Middle Cerebral Artery Occlusion Allowing Reperfusion via Common Carotid Artery Repair in Mice

The ischemic stroke is a major cause of adult long-term disability and death worldwide. The current treatments available are limited, with only tissue plasminogen activator (tPA) as an approved drug treatment to target ischemic strokes. Current research in the field of ischemic stroke focuses on better understanding the pathophysiology of stroke, to develop and investigate novel pharmaceutical targets. Reliable experimental stroke models are crucial for the progression of potential treatments. The middle cerebral artery occlusion (MCAO) model is clinically relevant and the most frequently used surgical model of ischemic stroke in rodents. However, the outcomes of this model, such as lesion volume, are associated with high levels of variability, particularly in mice. The alternative MCAO model described here allows the reperfusion of the common carotid artery (CCA) and the increased perfusion of the middle cerebral artery (MCA) territory, using a tissue pad with fibrinogen-based sealant to repair the vessel, and the improved welfare of the mice by avoiding external carotid artery (ECA) ligation. This reduces the reliance on the Circle of Willis, which is known to be highly anatomically variable in mice. Representative data show that using this alternative surgical approach decreases the variability in lesion volumes between the traditional MCAO approach and the alternative approach described here.

Intraluminal filament occlusion of the middle cerebral artery is the most frequently used in vivo model of experimental stroke in rodents. An alternative surgical approach to allow common carotid artery repair is performed here, which allows the reperfusion of the common carotid artery and a full reperfusion to the middle cerebral artery territory.

Oclusión de la arteria Cerebral Media permitiendo la reperfusión mediante la reparación de la arteria carótida común en ratones

The ischemic stroke is a major cause of adult long-term disability and death worldwide. The current treatments available are limited, with only tissue ...